LOCUS       sequence-213276-        6863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  translational elongation factor Tu.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6863)
  AUTHORS   Villarreal F, Contreras-Llano LE, Chavez M, Ding Y, Fan J, Pan T, 
            Tan C
  TITLE     Synthetic microbial consortia enable rapid assembly of pure 
            translation machinery.
  JOURNAL   Nat Chem Biol. 2018 Jan;14(1):29-35. doi: 10.1038/nchembio.2514. 
            Epub 2017 Nov 13.
  PUBMED    29131146
REFERENCE   2  (bases 1 to 6863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nchembio.2514"; journalName: "Nat Chem Biol"; date: 
            "2018-01"; volume: "14"; issue: "1"; pages: "29-35"
COMMENT     Sequence Label: B1-4
FEATURES             Location/Qualifiers
     source          1..6863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     171..190
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     274..293
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        274..292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             332..354
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1546..1563
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1637..1655)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1651..1698
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1873..1878,1879..1895,1896..1901))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1928..1946)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2014..2118
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2119..2187,2188..2979)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2337..2356)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3150..3738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3639..3658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3892..3909
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3924..4063
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3989..4008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4149..4171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4165..4356)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5668..6750)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6712..6731
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6751..6828)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
       61 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
      121 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
      181 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
      241 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattga
      301 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
      361 atgcattcta aagaaaaatt tgaacgtaca aaaccgcacg ttaacgttgg tactatcggc
      421 cacgttgacc acggtaaaac tactctgacc gctgcaatca ccaccgtact ggctaaaacc
      481 tacggcggtg ctgctcgtgc attcgaccag atcgataacg cgccggaaga aaaagctcgt
      541 ggtatcacca tcaacacttc tcacgttgaa tacgacaccc cgacccgtca ctacgcacac
      601 gtagactgcc cggggcacgc cgactatgtt aaaaacatga tcaccggtgc tgctcagatg
      661 gacggcgcga tcctggtagt tgctgcgact gacggcccga tgccgcagac tcgtgagcac
      721 atcctgctgg gtcgtcaggt aggcgttccg tacatcatcg tgttcctgaa caaatgcgac
      781 atggttgatg acgaagagct gctggaactg gttgaaatgg aagttcgtga acttctgtct
      841 cagtacgact tcccgggcga cgacactccg atcgttcgtg gttctgctct gaaagcgctg
      901 gaaggcgacg cagagtggga agcgaaaatc ctggaactgg ctggcttcct ggattcttat
      961 attccggaac cagagcgtgc gattgacaag ccgttcctgc tgccgatcga agacgtattc
     1021 tccatctccg gtcgtggtac cgttgttacc ggtcgtgtag aacgcggtat catcaaagtt
     1081 ggtgaagaag ttgaaatcgt tggtatcaaa gagactcaga agtctacctg tactggcgtt
     1141 gaaatgttcc gcaaactgct ggacgaaggc cgtgctggtg agaacgtagg tgttctgctg
     1201 cgtggtatca aacgtgaaga aatcgaacgt ggtcaggtac tggctaagcc gggcaccatc
     1261 aagccgcaca ccaagttcga atctgaagtg tacattctgt ccaaagatga aggcggccgt
     1321 catactccgt tcttcaaagg ctaccgtccg cagttctact tccgtactac tgacgtgact
     1381 ggtaccatcg aactgccgga aggcgtagag atggtaatgc cgggcgacaa catcaaaatg
     1441 gttgttaccc tgatccaccc gatcgcgatg gacgacggtc tgcgtttcgc aatccgtgaa
     1501 ggcggccgta ccgttggcgc gggcgttgtt gctaaagttc tgggccatca tcatcaccat
     1561 cattaaagca gaggccatat gctcgaggat ccggctgcta acaaagcccg aaaggaagct
     1621 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     1681 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatcccgc aagaggcccg
     1741 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     1801 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     1861 ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga
     1921 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1981 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2041 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2101 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2161 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2221 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2281 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2341 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     2401 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2461 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2521 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2581 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2641 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     2701 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2761 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2821 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2881 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2941 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3001 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3061 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3121 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3181 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3241 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3301 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3361 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3421 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3481 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3541 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3601 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3661 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3721 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3781 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3841 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3901 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3961 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4021 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     4081 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4141 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4201 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     4261 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     4321 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     4381 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     4441 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     4501 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     4561 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     4621 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     4681 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     4741 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     4801 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg
     4861 gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac
     4921 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
     4981 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
     5041 aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc
     5101 gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga
     5161 gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg
     5221 gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc
     5281 atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg
     5341 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     5401 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     5461 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     5521 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     5581 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     5641 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5701 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5761 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     5821 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     5881 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     5941 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     6001 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     6061 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     6121 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     6181 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     6241 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     6301 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     6361 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     6421 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     6481 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     6541 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     6601 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     6661 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     6721 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     6781 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     6841 gctctccctt atgcgactcc tgc
//
